mCD20-腺病毒穿梭质粒pDC315的构建

Construction of recombinant adenovirus shuttle plasmid with mCD20

目的:构建腺病毒穿梭质粒pDC315-mCD20重组质粒,为探讨编码mCD20基因的腺病毒载体做好准备,为CD20基因治疗淋巴瘤的研究奠定基础。方法:采用RT-PCR方法将CD20基因克隆出来,通过分子生物学技术构建过渡质粒pcDNA3.1-mCD20质粒,用PCR、酶切进行鉴定后,将该基因cDNA片段插入腺病毒穿梭质粒pDC315中,转化至高效感受态DH5a细菌细胞中进行扩增,提取质粒,获得重组质粒腺病毒穿梭质粒pDC315-mCD20。结果:经分析鉴定,重组质粒中含有mCD20基因,达到实验预期目的。结论:成功构建表达mCD20基因的重组质粒腺病毒穿梭质粒pDC315-mCD20,有助于后续研究构建腺病毒载体AdmCD20。

Objective To construct recombinant adenovirus shuttle plasmid PDC315-mCD20. Methods Mouse CD20 gene was synthesized with reverse transcription polymerase chain reaction （RT-PCR）, and then was cloned into pcDNA3.1 plasmid. After identification with restriction enzyme analysis, PCR and nucleotide sequencing analysis, the PDC315-mCD20 plasmid was inserted into adenovirus shuttle plasmid, and then the recombinant plasmid were transformed into the cells of DH5a bacteria. After amplification and abstraction, the recombinant plasmid pDC315-mCD20 was obtaind. Results The recombinant plasmid was confirmed containing mCD20 gene. Conclusion We have constructed recombinant adenovirus shuttle plasmid with mCD20 successfully, which may be helpful to the next study about the construction of the adenovirus mCD20.